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#f30;"></span></p><p><span>產(chǎn)品關(guān)鍵詞：</span>北京熒光定量PCR技術(shù)服務(wù),熒光定量PCR</p><p><span>***更新：</span>2020-08-22 14:16:12</p><p><span id="click" >瀏覽次數(shù)：0次</span></p><a rel="nofollow" class="button">聯(lián)系我們</a></div></div><div id="tifug87" class="rightbox"><h3>公司基本資料信息</h3><p style="border:0;"><a rel="nofollow">上海朝瑞生物科技有限公司</a></p><p><span>聯(lián)系人：</span>仲先生</p><p><span>郵箱: </span>scienceladder@163.com</p><p><span>電話: </span>18621867065</p><p><span>傳真: </span>021_</p><p><span>網(wǎng)址: </span></p><p><span>手機(jī): </span>021-54133071</p><p><span>地址: </span>上海市松江區(qū)廣富林路697號(hào)昂立大廈2113室</p><p>[當(dāng)前離線] <a rel="nofollow"> [加為商友] </a><a rel="nofollow"> [發(fā)送信件] </a></p></div></div><div id="r7prykr" class="page-proshow02"><p class="title">詳細(xì)說(shuō)明</p><div id="f8jqsu8" class="box"><p><p> 管家基因反應(yīng)體系：序號(hào) 反應(yīng)物劑量 1 SYBR Green 1 染料 10μl 2 內(nèi)參照上游引物F 0.5μl 3 內(nèi)參照下游引物R 0.5μl 4 dNTP 0.5μl 5 Taq酶 1μl 6 待測(cè)樣品cDNA 5μl 7 ddH2O 32.5μl 8 總體積 50μl 輕彈管底將溶液混合，6000rpm短暫離心，北京熒光定量PCR技術(shù)服務(wù)。 </p> <p> 制備好的陽(yáng)性標(biāo)準(zhǔn)品和檢測(cè)樣本同時(shí)上機(jī)，反應(yīng)條件為：93℃2分鐘，然后93℃ 1分鐘，55℃ 2分鐘，共40個(gè)循環(huán)。針對(duì)每一需要測(cè)量的基因，選擇一確定表達(dá)該基因的cDNA模板進(jìn)行PCR反應(yīng)。 </p> <p> 反應(yīng)體系：序號(hào) 反應(yīng)物劑量 1 10×PCR緩沖液 2.5 ul 2 MgCl2溶液 1，北京熒光定量PCR技術(shù)服務(wù).5 ul 3 上游引物F 0.5 ul 4 下游引物R 0.5 ul 5 dNTP混合液 3 ul 6 Taq聚合酶 1 ul 7 cDNA 1 ul 8 加水至總體積為 25ul 輕彈管底將溶液混合，6000rpm短暫離心。 </p> <p> 35個(gè)PCR循環(huán)(94℃1分鐘;55℃1分鐘;72℃1分鐘); 72℃延伸5分鐘。 </p> <p> PCR產(chǎn)物與 DNA Ladder在2%瓊脂糖凝膠電泳，溴化乙錠染色，檢測(cè)PCR產(chǎn)物是否為單一特異性擴(kuò)增條帶。 </p> <p> 將PCR產(chǎn)物進(jìn)行10倍梯度稀釋: 設(shè)定PCR產(chǎn)物濃度為1×1010，依次稀釋至109、108、107、106，北京熒光定量PCR技術(shù)服務(wù)、105、104幾個(gè)濃度梯度。所有cDNA樣品分別配置實(shí)時(shí)定量 PCR反應(yīng)體系。北京熒光定量PCR技術(shù)服務(wù)</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="北京熒光定量PCR技術(shù)服務(wù),熒光定量PCR" src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253046152344/6852880.png"></div><p style="text-indent:25px">溫度設(shè)置：在設(shè)置熒光 PCR 程序時(shí)，要仔細(xì)核對(duì)程序中的目標(biāo)溫度、恒溫時(shí)間和循環(huán)次數(shù)等參數(shù)，參數(shù)設(shè)置不正確將直接影響 PCR 檢測(cè)結(jié)果。延伸過(guò)程中要設(shè)置讀取收集熒光信號(hào)，如不設(shè)置收集熒光，將無(wú)法保存試驗(yàn)數(shù)據(jù)，無(wú)法判定檢測(cè)結(jié)果，造成試驗(yàn)失敗操作規(guī)范：在對(duì)樣品進(jìn)行核酸提取和實(shí)時(shí)熒光擴(kuò)增時(shí)要規(guī)范操作，防止樣品的交叉污染、酶的降解、假陽(yáng)性的出現(xiàn) 離心管、***頭和 PCR 管均需要高壓滅菌或使用商品化的無(wú) 污染的離心管、***頭和PCR 管。核酸提取結(jié)束后應(yīng)立即進(jìn)行儀器擴(kuò)增，提取的核酸不可長(zhǎng)時(shí)間置于室溫，易受空氣中酶的降解，短期可保存于－20 ℃冰箱，長(zhǎng)期應(yīng)保存于－70 ℃冰箱江蘇lncRNA熒光定量PCR課題承包</p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="北京熒光定量PCR技術(shù)服務(wù),熒光定量PCR" src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253046357422/4616552.png"></div><p> PCR檢測(cè)方法在臨床醫(yī)學(xué)及實(shí)驗(yàn)室檢驗(yàn)中**有價(jià)值的應(yīng)用領(lǐng)域就是對(duì)***性疾病的診斷。理論上，只要樣本有一個(gè)病原體存在，PCR方法就可以檢測(cè)到。該方法對(duì)病原體的檢測(cè)解決了免疫學(xué)檢測(cè)的“窗口期”問(wèn)題，常用于疾病的早期診斷，可判斷疾病是否處于隱性或亞臨床狀態(tài)。 </p> <p> 大量的熒光定量PCR研究資料表明，病原體數(shù)量與***性疾病病情的輕重程度、傳染性及***效果均有相關(guān)性。因此，通過(guò)熒光定量PCR方法得到的檢測(cè)結(jié)果，一定程度上可以準(zhǔn)確反映疾病***的情況。 </p><p> Scienceladder是讓知識(shí)、技術(shù)、專長(zhǎng)、成果、資源流淌起來(lái)的平臺(tái)！ Scienceladder是聚天下技術(shù)服務(wù)天下的平臺(tái)！ Scienceladder是你的知識(shí)、技術(shù)、專長(zhǎng)、成果實(shí)現(xiàn)價(jià)值轉(zhuǎn)化的平臺(tái)! 發(fā)布你的技術(shù)、成果、專長(zhǎng)，資源讓需求者得到幫助！發(fā)布你的需求，讓供給者幫助你！為科學(xué)做點(diǎn)事、為科學(xué)家做點(diǎn)事、為技術(shù)推廣做點(diǎn)事！ </p> <p> 歡迎來(lái)到上海朝瑞生物科技有限公司網(wǎng)站，聯(lián)系人是仲先生。 </p> <p> 主要經(jīng)營(yíng)科研技術(shù)服務(wù)|應(yīng)用技術(shù)服務(wù)|科研成果轉(zhuǎn)化|科學(xué)產(chǎn)品銷售。 </p> <p> <br /> </p><div style="width:100%;text-align: center;"><img style="max-width: 640px; max-height: 320px;" alt="北京熒光定量PCR技術(shù)服務(wù),熒光定量PCR" src="http://tyunfile.71360.com/UploadFile/shchaorui/637116253057744141/7327894.png"></div><p> 熒光定量PCR(Flurogenic Quantitative Polymerase ChainReaction，F(xiàn)Q-PCR；real-time quantitative PCR, RT-qPCR or qPCR)，是1996 年由美國(guó)Applied Biosystems 公司推出的一種新定量試驗(yàn)技術(shù)，它是通過(guò)熒光染料或熒光標(biāo)記的特異性探針，標(biāo)記**PCR產(chǎn)物進(jìn)行實(shí)時(shí)監(jiān)測(cè)反應(yīng)，利用與之相適應(yīng)的軟件對(duì)產(chǎn)物進(jìn)行分析，計(jì)算待測(cè)樣品模板的初始濃度 [1] 。 </p> <p> 熒光域值(threshold)的設(shè)定：PCR反應(yīng)的前15個(gè)循環(huán)的熒光信號(hào)作為熒光本底信號(hào)，熒光域值的缺省設(shè)置是3～15個(gè)循環(huán)的熒光信號(hào)標(biāo)準(zhǔn)偏差的10倍北京熒光定量PCR技術(shù)服務(wù)</p><p style="text-indent:25px">北京熒光定量PCR技術(shù)服務(wù)</p><p style="text-indent:25px">比較Ct值的相對(duì)定量法：此方法是將待測(cè)靶基因片段和內(nèi)參照基因片段同時(shí)擴(kuò)增，可以在兩個(gè)反應(yīng)管中或者一個(gè)反應(yīng)管中進(jìn)行擴(kuò)增反應(yīng)，并且就兩者的ct值之差進(jìn)行測(cè)量。在采用此方法過(guò)程中需要用數(shù)學(xué)公式進(jìn)行相對(duì)量的計(jì)算，首先要假設(shè)每個(gè)循環(huán)產(chǎn)物增加一倍時(shí)PCR反應(yīng)**期得到Ct值對(duì)起始模板的量一個(gè)循環(huán)的估算和起始模板數(shù)兩倍相當(dāng)。這種方法的前提是靶基因和內(nèi)源控制物的擴(kuò)增效率基本一致，所以在應(yīng)用過(guò)程中效率的偏移會(huì)對(duì)實(shí)際拷貝數(shù)估計(jì)產(chǎn)生一定影響，因此，為了保證目的基因和內(nèi)參基因的擴(kuò)增效率相等應(yīng)當(dāng)加強(qiáng)對(duì)反應(yīng)體系的優(yōu)化，這也是該方法應(yīng)用的難點(diǎn)之一北京熒光定量PCR技術(shù)服務(wù)</p><p style="text-indent:25px">上海朝瑞生物科技有限公司屬于化工的高新企業(yè)，技術(shù)力量雄厚。上海朝瑞科技是一家有限責(zé)任公司企業(yè)，一直“以人為本，服務(wù)于社會(huì)”的經(jīng)營(yíng)理念;“質(zhì)量高速，誠(chéng)守信譽(yù)，持續(xù)發(fā)展”的質(zhì)量方針。公司業(yè)務(wù)涵蓋[ "科研技術(shù)服務(wù)", "應(yīng)用技術(shù)服務(wù)", "科研成果轉(zhuǎn)化", "科學(xué)產(chǎn)品銷售" ]，價(jià)格合理，品質(zhì)有保證，深受廣大客戶的歡迎。上海朝瑞科技自成立以來(lái)，一直堅(jiān)持走正規(guī)化、專業(yè)化路線，得到了廣大客戶及社會(huì)各界的普遍認(rèn)可與大力支持。</p></p><br /><p>文章來(lái)源地址: 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